Biochemical and functional properties of vesicles from planktonic and biofilm phenotypes of Limosilactobacillus reuteri DSM 17938
Abstract Limosilactobacillus reuteri DSM 17938 is among the world’s most studied probiotic strains and has been shown to provide several health benefits for the host. We have previously shown that the cell-free supernatant of L. reuteri DSM 17938 possesses antimicrobial activity and contains several...
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Nature Portfolio
2025-05-01
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| Online Access: | https://doi.org/10.1038/s41598-025-03823-w |
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| author | Beatrice Marinacci Chiara D’Ambrosio Irene Vitale Antonella Di Sotto Francesco Cairone Mattia Spano Simone Carradori Andrea Scaloni Marco Gullì Valentina Puca Santolo Francati Monica Matuozzo Ludwig Ermann Lundberg Gianfranco Grompone Stefan Roos Rossella Grande |
| author_facet | Beatrice Marinacci Chiara D’Ambrosio Irene Vitale Antonella Di Sotto Francesco Cairone Mattia Spano Simone Carradori Andrea Scaloni Marco Gullì Valentina Puca Santolo Francati Monica Matuozzo Ludwig Ermann Lundberg Gianfranco Grompone Stefan Roos Rossella Grande |
| author_sort | Beatrice Marinacci |
| collection | DOAJ |
| description | Abstract Limosilactobacillus reuteri DSM 17938 is among the world’s most studied probiotic strains and has been shown to provide several health benefits for the host. We have previously shown that the cell-free supernatant of L. reuteri DSM 17938 possesses antimicrobial activity and contains several bioactive compounds. Furthermore, the strain was shown to be a biofilm producer that releases both planktonic and biofilm Membrane Vesicles (MVs). In this study, membrane vesicles isolated from planktonic (pMVs) and biofilm (bMVs) phenotypes were comparatively investigated for their toxicity, ability to kill cancer as well as non-cancer cell lines and modulate phagocytosis in murine macrophages. Neither pMVs nor bMVs showed any in vivo toxicity in a Galleria mellonella model, and weakly affected cancer and noncancerous cell viability after both short- and long-term treatments. However, they were able to affect phagocytosis in lipopolysaccharide challenged RAW 264.7 macrophages, suggesting possible immunomodulatory properties. NMR-based metabolomic analysis of pMVs and bMVs identified and quantified engulfed compounds, mainly organic acids and amino acids, with lactate being the most abundant molecule in both vesicle types. bMVs contained higher concentrations of all measured metabolites compared to pMVs. Proteomic analysis of pMVs and bMVs described equivalent protein cargos, emphasizing quantitative compositional differences that presumably reflect the physiological state of each parent bacterial phenotype. Through the assignment of molecules possibly acting as mediators of immune/inflammatory responses in the host and/or modulating known beneficial effects of L. reuteri, important signaling functions of these vesicles were suggested. Finally, storage stability of MVs up to four weeks was established. |
| format | Article |
| id | doaj-art-b6f4651f02ff4ec29015a3d075d405cc |
| institution | OA Journals |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-b6f4651f02ff4ec29015a3d075d405cc2025-08-20T02:03:34ZengNature PortfolioScientific Reports2045-23222025-05-0115112110.1038/s41598-025-03823-wBiochemical and functional properties of vesicles from planktonic and biofilm phenotypes of Limosilactobacillus reuteri DSM 17938Beatrice Marinacci0Chiara D’Ambrosio1Irene Vitale2Antonella Di Sotto3Francesco Cairone4Mattia Spano5Simone Carradori6Andrea Scaloni7Marco Gullì8Valentina Puca9Santolo Francati10Monica Matuozzo11Ludwig Ermann Lundberg12Gianfranco Grompone13Stefan Roos14Rossella Grande15Department of Pharmacy, “G. d’Annunzio” University of Chieti-PescaraProteomics, Metabolomics and Mass Spectrometry Laboratory, ISPAAM- National Research CouncilDepartment of Pharmacy, “G. d’Annunzio” University of Chieti-PescaraDepartment of Physiology and Pharmacology “V. Erspamer”, Sapienza University of RomeDepartment of Chemistry and Technology of Drugs, Sapienza University of RomeDepartment of Chemistry and Technology of Drugs, Sapienza University of RomeDepartment of Pharmacy, “G. d’Annunzio” University of Chieti-PescaraProteomics, Metabolomics and Mass Spectrometry Laboratory, ISPAAM- National Research CouncilDepartment of Physiology and Pharmacology “V. Erspamer”, Sapienza University of RomeDepartment of Pharmacy, “G. d’Annunzio” University of Chieti-PescaraDepartment of Agricultural and Food Sciences (DISTAL), University of BolognaProteomics, Metabolomics and Mass Spectrometry Laboratory, ISPAAM- National Research CouncilDepartment of Molecular Sciences, Uppsala Biocenter, Swedish University of Agricultural SciencesBioGaia ABDepartment of Molecular Sciences, Uppsala Biocenter, Swedish University of Agricultural SciencesDepartment of Pharmacy, “G. d’Annunzio” University of Chieti-PescaraAbstract Limosilactobacillus reuteri DSM 17938 is among the world’s most studied probiotic strains and has been shown to provide several health benefits for the host. We have previously shown that the cell-free supernatant of L. reuteri DSM 17938 possesses antimicrobial activity and contains several bioactive compounds. Furthermore, the strain was shown to be a biofilm producer that releases both planktonic and biofilm Membrane Vesicles (MVs). In this study, membrane vesicles isolated from planktonic (pMVs) and biofilm (bMVs) phenotypes were comparatively investigated for their toxicity, ability to kill cancer as well as non-cancer cell lines and modulate phagocytosis in murine macrophages. Neither pMVs nor bMVs showed any in vivo toxicity in a Galleria mellonella model, and weakly affected cancer and noncancerous cell viability after both short- and long-term treatments. However, they were able to affect phagocytosis in lipopolysaccharide challenged RAW 264.7 macrophages, suggesting possible immunomodulatory properties. NMR-based metabolomic analysis of pMVs and bMVs identified and quantified engulfed compounds, mainly organic acids and amino acids, with lactate being the most abundant molecule in both vesicle types. bMVs contained higher concentrations of all measured metabolites compared to pMVs. Proteomic analysis of pMVs and bMVs described equivalent protein cargos, emphasizing quantitative compositional differences that presumably reflect the physiological state of each parent bacterial phenotype. Through the assignment of molecules possibly acting as mediators of immune/inflammatory responses in the host and/or modulating known beneficial effects of L. reuteri, important signaling functions of these vesicles were suggested. Finally, storage stability of MVs up to four weeks was established.https://doi.org/10.1038/s41598-025-03823-wLactobacillus reuteriLimosilactobacillus reuteriExtracellular membrane vesiclesProteomicsMetabolomicsCytotoxicity |
| spellingShingle | Beatrice Marinacci Chiara D’Ambrosio Irene Vitale Antonella Di Sotto Francesco Cairone Mattia Spano Simone Carradori Andrea Scaloni Marco Gullì Valentina Puca Santolo Francati Monica Matuozzo Ludwig Ermann Lundberg Gianfranco Grompone Stefan Roos Rossella Grande Biochemical and functional properties of vesicles from planktonic and biofilm phenotypes of Limosilactobacillus reuteri DSM 17938 Scientific Reports Lactobacillus reuteri Limosilactobacillus reuteri Extracellular membrane vesicles Proteomics Metabolomics Cytotoxicity |
| title | Biochemical and functional properties of vesicles from planktonic and biofilm phenotypes of Limosilactobacillus reuteri DSM 17938 |
| title_full | Biochemical and functional properties of vesicles from planktonic and biofilm phenotypes of Limosilactobacillus reuteri DSM 17938 |
| title_fullStr | Biochemical and functional properties of vesicles from planktonic and biofilm phenotypes of Limosilactobacillus reuteri DSM 17938 |
| title_full_unstemmed | Biochemical and functional properties of vesicles from planktonic and biofilm phenotypes of Limosilactobacillus reuteri DSM 17938 |
| title_short | Biochemical and functional properties of vesicles from planktonic and biofilm phenotypes of Limosilactobacillus reuteri DSM 17938 |
| title_sort | biochemical and functional properties of vesicles from planktonic and biofilm phenotypes of limosilactobacillus reuteri dsm 17938 |
| topic | Lactobacillus reuteri Limosilactobacillus reuteri Extracellular membrane vesicles Proteomics Metabolomics Cytotoxicity |
| url | https://doi.org/10.1038/s41598-025-03823-w |
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